筛选普氏原羚粪便DNA中微卫星引物 并应用于个体识别

为了更好地保护极度濒危的普氏原羚物种，选择非损伤性样品-粪便作为研究材料，选用10对非洲糜羚微卫星引物和10对绵羊微卫星引物作为筛选普氏原羚基因组DNA微卫星位点的引物。通过非变性聚丙烯酰胺凝胶电泳检测微卫星PCR的扩增产物，结果发现20对引物中有8对引物在普氏原羚基因组DNA中扩增出了多态性位点。通过等位基因数目和等位基因频率对这8个位点的基因杂合度、多态性信息含量、有效等位基因数进行了计算，结果发现这8个位点在39个普氏原羚粪便样品中的基因杂合度介于0.71~0.84，平均杂合度为0.78；多态性信息含量介于0.79~0.66，平均多态信息含量为0.73；有效等位基因数介于3.40~6.08，平均有效等位基因为5.98，这表明所筛选到的8个微卫星基因座在研究普氏原羚粪便样品中均为中高度多态性基因座，具有比较明显的遗传变异，完全适合普氏原羚各种分子遗传分析。因此试验应用这8对多态性引物对39个粪便样品的个体进行识别，发现这39个粪便样品来自35个不同的个体。

Screening Microsatellite Primers and their Application for Individual Identification in Faecal DNA from Procapra praprzewalskii

To protect Procapra praprzewalskii appointed as CR (Critically Endangered), twenty pairs of microsatellite primers from Granti’s gazelle and sheep separately were used to screen microsatellite locus in faecal DNA from Procapra praprzewalskii, eight polymorphism loci was found. The PCR amplified products of microsatellite loci were detected y non-denatured polyacrylamide gel electrophoresis. Gene heterozygosity (He), polymorphism information content (PIC) and effective number of allele (Ne) from the eight loci were calculated by allele numbers and allelic frequency. The heterozygosity (He) ranged from0.71 to 0.84, the average was 0.78; the PIC were from 0.79 to 0.66, their average was 0.73; and Ne were from 3.40 to 6.08, the average was 5.98;which showed the eight microsatellite loci had high or medium polymorphism in the Procapra raprzewalskii and had high genetic diversity. So the 8 loci were use to identify Individual, the result was 35 individuals from 39 faecal samples from Procapra praprzewalskii.